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结肠炎抗原特异性模型中淋巴细胞蛋白质谱的改变:一项比较蛋白质组学研究。

Altered protein profile of lymphocytes in an antigen-specific model of colitis: a comparative proteomic study.

作者信息

Liu B-G, Cao Y-B, Cao Y-Y, Zhang J-D, An M-M, Wang Y, Gao P-H, Yan L, Xu Y, Jiang Y-Y

机构信息

Department of Pharmacology, School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai, 200433, China.

出版信息

Inflamm Res. 2007 Sep;56(9):377-84. doi: 10.1007/s00011-007-7035-0.

Abstract

OBJECTIVE

Lymphocytes are deeply involved in the initiation and perpetuation of inflammatory response in inflammatory bowel disease (IBD) and lymphocyte-derived proteins are associated with the pathogenesis of the disease. The aim of this study was to identify the altered protein profiles of lymphocytes from rats with colitis.

METHODS

Colitis models were induced by colonic administration of trinitrobenzene sulfonic acid (TNBS) in 50% ethanol in male SD rats. Seven days after administration of TNBS/ethanol, lymphocytes were harvested from mesenteric lymph nodes (MLNs) and proteins were extracted. Two-dimensional polyacrylamide gel electrophoresis and PDQuest 2D-image-analysis software were used to display and analyze the protein spots. The differentially-expressed proteins were identified by tryptic in-gel digestion and mass spectrometry. Real-time RT-PCR was used for selected transcripts to validate the findings of the proteomics analysis.

RESULTS

A total of 1,100 protein spots including 26 proteins with at least a two-fold difference in abundance between colitis and control groups were identified. Among all the detected spots, 17 were up-regulated and 9 were down-regulated. It was found that the altered proteins included the regulators of the cell cycle and cell proliferation, signal transduction factors, inflammatory factors, apoptosis-related proteins and metabolic enzymes.

CONCLUSIONS

In lymphocytes of rats with TNBS-induced colitis, 26 altered proteins were identified. They involve inflammation, apoptosis, metabolism, and regulation of the cell cycle, cell proliferation, and signal transduction.

摘要

目的

淋巴细胞深度参与炎症性肠病(IBD)炎症反应的起始和持续,且淋巴细胞衍生蛋白与该疾病的发病机制相关。本研究旨在鉴定结肠炎大鼠淋巴细胞中改变的蛋白质谱。

方法

通过在雄性SD大鼠中结肠内给予三硝基苯磺酸(TNBS)于50%乙醇中诱导结肠炎模型。给予TNBS/乙醇7天后,从肠系膜淋巴结(MLN)中收获淋巴细胞并提取蛋白质。使用二维聚丙烯酰胺凝胶电泳和PDQuest 2D图像分析软件来展示和分析蛋白质斑点。通过胰蛋白酶胶内消化和质谱鉴定差异表达的蛋白质。对选定的转录本使用实时RT-PCR来验证蛋白质组学分析的结果。

结果

共鉴定出1100个蛋白质斑点,其中包括26种在结肠炎组和对照组之间丰度至少有两倍差异的蛋白质。在所有检测到的斑点中,17个上调,9个下调。发现改变的蛋白质包括细胞周期和细胞增殖的调节因子、信号转导因子、炎症因子、凋亡相关蛋白和代谢酶。

结论

在TNBS诱导的结肠炎大鼠的淋巴细胞中,鉴定出26种改变的蛋白质。它们涉及炎症、凋亡、代谢以及细胞周期、细胞增殖和信号转导的调节。

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